Chen Xiaojuan, Jin Guoying, Luo Hong, Zhou Lifei
Zhejiang Cancer Hospital, Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou, Zhejiang, China.
Anticancer Drugs. 2023 Jul 1;34(6):725-734. doi: 10.1097/CAD.0000000000001460. Epub 2023 Jan 24.
The aim of this study is to reveal the mechanism of Gubenyiliu II (GYII) inhibiting autophagy in breast cancer and the effect of its disassembled prescriptions, Quxie (QX) and Fuzheng (FZ), which cause autophagy difference on tumor growth. After a breast cancer in situ tumor model was established, mice were randomly distributed into different groups: model, GYII, QX, FZ and tamoxifen groups, and treated correspondingly. Then, the tumor volumes and weights were monitored. Immunohistochemistry detected the contents of microtubule-associated protein light chain 3 (LC3), phosphorylated phosphatidylinositol 3-kinase (p-PI3K), phosphorylated protein kinase B (p-AKT) and phosphorylated mammalian target of rapamycin (p-mTOR) in tumor tissues. Furthermore, 4T1 cells were administrated with the 20% contained serum. Cell proliferation, migration and invasion were measured using cell counting kit-8 and transwell assays. Electron microscopy and flow cytometry detected autophagy and apoptosis. The content of LC3 was measured by immunofluorescence. Western blot detected the protein levels of LC3, Beclin1, p-PI3K/PI3K, p-AKT/AKT and p-mTOR/mTOR in tumor tissues and 4T1 cells. GYII, QX and FZ treatment significantly reduced the tumor volumes and weights in breast cancer tumor-bearing mice. The cell proliferation, migration and invasion were restrained, and cell apoptosis and autophagy were promoted in GYII, QX and FZ groups. Moreover, GYII, QX and FZ increased the expression of LC3 in 4T1 cells and tumor tissues and decreased the phosphorylation levels of PI3K, AKT and mTOR in tumor tissues. The protein levels of LC3 and Beclin1 were upregulated, and p-PI3K/PI3K, p-AKT/AKT and p-mTOR/mTOR were downregulated in tumor tissues and 4T1 cells of treatment groups. Our study confirmed that GYII could treat breast cancer by restraining the PI3K/AKT/mTOR signaling pathway-mediated autophagy. While QX focuses on inhibiting tumor growth, FZ acts on inhibiting tumor metastasis.
本研究旨在揭示固本抑瘤Ⅱ号（GYII）抑制乳腺癌自噬的机制及其拆方祛邪（QX）和扶正（FZ）导致自噬差异对肿瘤生长的影响。建立乳腺癌原位肿瘤模型后，将小鼠随机分为不同组：模型组、GYII组、QX组、FZ组和他莫昔芬组，并进行相应治疗。然后，监测肿瘤体积和重量。免疫组织化学检测肿瘤组织中微管相关蛋白轻链3（LC3）、磷酸化磷脂酰肌醇3激酶（p-PI3K）、磷酸化蛋白激酶B（p-AKT）和磷酸化雷帕霉素靶蛋白（p-mTOR）的含量。此外，用含20%血清处理4T1细胞。使用细胞计数试剂盒-8和Transwell实验检测细胞增殖、迁移和侵袭。电子显微镜和流式细胞术检测自噬和凋亡。通过免疫荧光检测LC3的含量。蛋白质印迹法检测肿瘤组织和4T1细胞中LC3、Beclin1、p-PI3K/PI3K、p-AKT/AKT和p-mTOR/mTOR的蛋白水平。GYII、QX和FZ治疗显著降低了荷乳腺癌小鼠的肿瘤体积和重量。在GYII、QX和FZ组中，细胞增殖、迁移和侵袭受到抑制，细胞凋亡和自噬得到促进。此外，GYII、QX和FZ增加了4T1细胞和肿瘤组织中LC3的表达，并降低了肿瘤组织中PI3K、AKT和mTOR的磷酸化水平。治疗组肿瘤组织和4T1细胞中LC3和Beclin1的蛋白水平上调，p-PI3K/PI3K、p-AKT/AKT和p-mTOR/mTOR下调。我们的研究证实，GYII可通过抑制PI3K/AKT/mTOR信号通路介导的自噬来治疗乳腺癌。QX侧重于抑制肿瘤生长，而FZ则作用于抑制肿瘤转移。
